Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

Characterization of allergoids from ovalbumin in vitro and in vivo

Several in vivo and in vitro methods for monitoring immunological properties of two allergoids obtained by formaldehyde treatment of ovalbumin (OA) were developed. The calculated molecular weight of allergoids was 80 kD (OA-F1) and 165 kD (OA-F2), respectively. The allergenic activity in vitro of allergoids in mast-cell histamine release assay was 1000 times lower than of OA. Both allergoids showed reduced ability to induce passive cutaneous anaphylaxis in the Sprague-Dawley rats or systemic anaphylaxis in Dunkin-Harley guinea-pigs. The ability of OA and allergoids to bind to the OA-specific IgE antibodies was measured in vivo by the inhibition of passive cutaneous anaphylaxis (PCA-inhibition). Allergoid binding to IgE was 51-66% lower than the native allergen. Moreover, the avidity of OA-specific IgG antibodies, measured by ELISA-inhibition, for allergoids and allergen was of the same order. Allergoids induced a different pattern of humoral immune response from that, induced by the native allergen. Thus, after immunization of BALB/c mouse, both allergoids induced a higher production of IgG and a lower production of IgE than OA, only OA-F2 induced a lower production of IgG1. The differences in the IgA response to the immunogens was not significant. Delayed hypersensitivity studies in the BALB/c mouse showed that allergoids were 5- to 12-times less effective in inducing a cell-mediated immune response than OA. The present study provides a battery of immunological methods for preclinical testing of modified allergens.     

Deformation and fracture of glass bead/ CTBN-rubber/epoxy composites

Carboxyl-terminated butadiene-acrylonitrile-rubber decreases modulus and yield stress of the studied epoxy but increases fracture toughness. The addition of glass bead compensates for the loss in modulus but has little effect on yield stress. However, it significantly contributes to the fracture toughness by providing additional mechanisms for toughening of both the unmodified and rubber-modified epoxy. For the toughened epoxies studied, fracture surfaces gave only limited information on fracture mechanisms since significant energy absorption also occurs in the material below the fracture surface. Suggestions for suitable material compositions for fiber composite matrices are given.     

Cold forging and chemical heat treatment of the casing of the internal joint for VAZ cars

The technological process of cold forging applied for the first time in the production of the casing of the internal joint with races is described. The process operations of cold forging and the annealing and carburizing regimes for this part me described.     

Afterburner [network-independent card for protocols]

Many current implementations of protocols such as the Transmission Control Protocol/Internet Protocol (TCP/IP) are inefficient because data are often accessed more frequently than necessary. Three techniques that reduce the need for memory bandwidth are proposed. The techniques are copy-on-write, page remapping, and single-copy. Afterburner, a network-independent card that provides the services that are necessary for a single-copy protocol stack, is described. The card has 1 MByte of local buffers and provides a simple interface to a variety of network link adapters, including HIPPI and asynchronous transfer mode (ATM). Afterburner can support transfers to and from the link adapter card at rates up to 1 Gbit/s. An implementation of TCP/IP that uses the features provided by Afterburner to reduce the movement of data to a single copy is discussed. Measurements of the end-to-end performance of Afterburner and the single-copy implementation of TCP/IP are presented     

Functional Properties and Food Applications of Rapeseed Protein Concentrate

Rapeseed protein concentrate (RC), prepared with 2% hexameta-phosphate, was tested for its functionality and performance in some foods. The RC had good nitrogen solubility, fat absorption, emulsification, and whipping capacities but poor water absorption and gelling properties. It increased the emulsion stability, and protein but lowered the fat content of wieners. It also increased the cooking yield, reduced the shrinkage and tenderized meat patties. Results were similar to soybean isolate except for the poorer color and flavor. The cooking yield of RC supplemented wieners was less than the all-meat control and soybean-supplemented wieners. A 9% RC dispersion mixed with an equal volume of eggwhite produced a meringue of comparable stability and texture to that of eggwhite alone.     

Interdisciplinary research: roles for self-organization

No generally accepted principles and guidelines currently exist to help engineers design local interaction mechanisms that result in a desired global behavior. However, several communities have developed ways of approaching this problem in the context of niched application areas. Because the ideas underlying these approaches are often obscured or underemphasized in technical papers, the authors review the role of self-organization in their work. They provide a better picture of the status of the emerging field of self-organizing systems or autonomic computing.